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Literature summary for 6.2.1.13 extracted from

  • Schmidt, M.; Schoenheit, P.
    Acetate formation in the photoheterotrophic bacterium Chloroflexus aurantiacus involves an archaeal type ADP-forming acetyl-CoA synthetase isoenzyme I (2013), FEMS Microbiol. Lett., 349, 171-179.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Chloroflexus aurantiacus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.024
-
CoA in 100 mM HEPES-KOH pH 7.5, 0.1 mM 5,5'-dithiobis(2-nitrobenzoic acid), 5 mM MgCl2, at 55°C Chloroflexus aurantiacus
0.037
-
acetyl-CoA in 100 mM HEPES-KOH pH 7.5, 0.1 mM 5,5'-dithiobis(2-nitrobenzoic acid), 5 mM MgCl2, at 55°C Chloroflexus aurantiacus
0.091
-
ADP in 100 mM HEPES-KOH pH 7.5, 0.1 mM 5,5'-dithiobis(2-nitrobenzoic acid), 5 mM MgCl2, at 55°C Chloroflexus aurantiacus
0.57
-
ATP in 100 mM HEPES-KOH pH 7.5, 0.1 mM 5,5'-dithiobis(2-nitrobenzoic acid), 5 mM MgCl2, at 55°C Chloroflexus aurantiacus
0.9
-
acetate in 100 mM HEPES-KOH pH 7.5, 0.1 mM 5,5'-dithiobis(2-nitrobenzoic acid), 5 mM MgCl2, at 55°C Chloroflexus aurantiacus
1
-
phosphate in 100 mM HEPES-KOH pH 7.5, 0.1 mM 5,5'-dithiobis(2-nitrobenzoic acid), 5 mM MgCl2, at 55°C Chloroflexus aurantiacus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ dependent on Mg2+, showing the highest activity at about 5 mM Chloroflexus aurantiacus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
75000
-
4 * 75000, native enzyme, SDS-PAGE Chloroflexus aurantiacus
77000
-
4 * 77000, recombinant enzyme, SDS-PAGE Chloroflexus aurantiacus
300000
-
native enzyme, gel filtration Chloroflexus aurantiacus
317000
-
recombinant enzyme, gel filtration Chloroflexus aurantiacus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP + phosphate + acetyl-CoA Chloroflexus aurantiacus
-
ATP + acetate + CoA
-
r
ADP + phosphate + acetyl-CoA Chloroflexus aurantiacus J-10-fl
-
ATP + acetate + CoA
-
r
ATP + acetate + CoA Chloroflexus aurantiacus
-
ADP + phosphate + acetyl-CoA
-
r
ATP + acetate + CoA Chloroflexus aurantiacus J-10-fl
-
ADP + phosphate + acetyl-CoA
-
r

Organism

Organism UniProt Comment Textmining
Chloroflexus aurantiacus
-
-
-
Chloroflexus aurantiacus J-10-fl
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA affinity chromatography, phenyl Sepharose column chromatography, Q-Sepharose column chromatography, and Superdex 200 gel filtration Chloroflexus aurantiacus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + phosphate + acetyl-CoA
-
Chloroflexus aurantiacus ATP + acetate + CoA
-
r
ADP + phosphate + acetyl-CoA
-
Chloroflexus aurantiacus J-10-fl ATP + acetate + CoA
-
r
ATP + 2-methylbutyrate + CoA 75% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + 2-methylbutyryl-CoA
-
?
ATP + acetate + CoA
-
Chloroflexus aurantiacus ADP + phosphate + acetyl-CoA
-
r
ATP + acetate + CoA 100% activity Chloroflexus aurantiacus ADP + phosphate + acetyl-CoA
-
r
ATP + acetate + CoA
-
Chloroflexus aurantiacus J-10-fl ADP + phosphate + acetyl-CoA
-
r
ATP + butyrate + CoA 98% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + butyryl-CoA
-
?
ATP + imidazole-4-acetate + CoA 17% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + imidazol-4-ylacetyl-CoA
-
?
ATP + isobutyrate + CoA 46% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + isobutyryl-CoA
-
?
ATP + isobutyrate + CoA 46% activity compared to acetate Chloroflexus aurantiacus J-10-fl ADP + phosphate + isobutyryl-CoA
-
?
ATP + isovalerate + CoA 65% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + isovaleryl-CoA
-
?
ATP + phenylacetate + CoA 10% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + phenylacetyl-CoA
-
?
ATP + propionate + CoA 100% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + propionyl-CoA
-
?
ATP + propionate + CoA 100% activity compared to acetate Chloroflexus aurantiacus J-10-fl ADP + phosphate + propionyl-CoA
-
?
ATP + thioglycolate + CoA 110% activity compared to acetate Chloroflexus aurantiacus ADP + phosphate + thioglycolyl-CoA
-
?
additional information 4-hydroxyphenylacetate, indol-3-acetate and succinate are not utilized as substrates Chloroflexus aurantiacus ?
-
?
additional information 4-hydroxyphenylacetate, indol-3-acetate and succinate are not utilized as substrates Chloroflexus aurantiacus J-10-fl ?
-
?

Subunits

Subunits Comment Organism
homotetramer 4 * 75000, native enzyme, SDS-PAGE Chloroflexus aurantiacus
homotetramer 4 * 77000, recombinant enzyme, SDS-PAGE Chloroflexus aurantiacus

Synonyms

Synonyms Comment Organism
ACD
-
Chloroflexus aurantiacus
Acetyl-CoA synthetase (ADP-forming)
-
Chloroflexus aurantiacus
ADP-forming acetyl-CoA synthetase isoenzyme I
-
Chloroflexus aurantiacus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
Chloroflexus aurantiacus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Chloroflexus aurantiacus

pH Range

pH Minimum pH Maximum Comment Organism
7.3 7.8 the pH optimum is at pH 7.5, with remaining activities of about 40% at pH 7.3 and 59% at pH 7.8 Chloroflexus aurantiacus

Cofactor

Cofactor Comment Organism Structure
ATP
-
Chloroflexus aurantiacus

Expression

Organism Comment Expression
Chloroflexus aurantiacus the enzyme is induced 10fold during growth on D-glucose up